Novel pregnadiene

ABSTRACT

17 Alpha ,21-DIMETHYL-19-NOR- Delta 4,9-PREGNADIENE-3,20-DIONE OF THE FORMULA   HAVING ANTIGONADO-HYPOPHYSIAL, PROGESTOMIMETIC AND PROGESTATIVE ACTIVITY AND ITS PREPARATION.

United States Patent 11 1 Warnant et al.

[ NOVEL PREGNADIENE [75] Inventors: Julien Warnant, Neuilly S/Seine;

Andre Farcilli, Rosny-sous-Bois, both of France [73] Assignee: Roussel Uclaf, Paris, France [22] Filed: Feb. 29, 1972 [21] Appl. No.: 230,507

I Related US. Application Data [62] Division of Ser. No. 112,387, Feb. 3, 1971, Pat. No.

[30] Foreign Application Priority Data Feb. 20, 1970 France 7006179 [52] U.S. Cl. 424/242 [51] Int. Cl A6lk 17/00 [58] Field of Search 424/242; 260/397.3

[56] References Cited UNlTED STATES PATENTS 3,086,027 4/1963 Perelman et al 260/397.3

[451 Sept. 25, 1973 3,277,122 10/1966 Alvarez 260/397.3 3,679,714 7/1972 Warnant et al. 260/397.3

Primary ExaminerShep K. Rose AttorneyHammond & Littell [57] ABSTRACT 1 701,2 1 -dirnethyl- 19-nor-A -pregnadiene-3 ,20-dione of the formula (Fug-CH3 C:()

having antigonado-hypophysial, progestomimetic and progestative activity and its preparation.

5 Claims, N0 Drawings NOVEL PREGNADIENE This is a division of Ser. No. 112,387, filed Feb. 3, 1971 now US. Pat. No. 3,679,714.

OBJECTS OF THE INVENTION It is an object of the invention to provide the novel product, 17a,21dimethyl-l9-nor-A -pregnadiene- 3,20-dione.

It is another object of the invention to provide a novel process for the preparation of 17a,21-dimethyl- 19-nor-N- -pregnadiene-3,20-dione.

It is another object of the invention to provide novel pharmaceutical compositions having antigonadohypophysial, progestomimetic and progestative activity.

It is a further object of the invention to provide ,a method of inducing antigonado-hypophysial, progestomimetic and progestative activity in warm blooded animals.

These and other objects and advantages of the invention will become obvious from the following description.

THE INVENTION The novel product of the invention is 1701,21- dimethyl-19-nor-A- -pregnadiene-3,20-dione and has interesting pharmacological properties, namely .antigonado-hypophysial activity, progestomimetic activity and progestative activity.

The novel process of the invention for the preparation of 17a,21-dimeth.yl-19-nor-A- -pregnadiene-6 21a3,20-dione comprises reacting 3-methoxy-19-nor- A" -pregnate-traene-20-one with an alkali metal and a methyl halide to simultaneously reduce the 16,17 double bond and methylate which results in the formation of 3-methoxy-17a-methyl-19-nor-A- pregnatriene-ZO-one, reducing the latter by the Djerassi method with an alkali metal in the presence ofammonia to form 3-methoxy-17a-methyl-19-nor-A pregnadiene-ZO-ol, hydrolyzing the latter with a dilute acid to obtain 17oi-methyl-19-nor-A "-pregnene-20-ol-3one, oxidizing the latter with an oxidizing acid agent to form 17a-methyl-l9-nor-A- pregnene-3,20-dione and successively brominating with bromine and dehydrobrominating with pyridine to form 17a-methyl-19-nor-A" -pregnadiene-3,20-dione and recovering from the crystallization mother liquors 1701,21dimethyl-l9-nor-A--pregnadiene-3, 20-dione.

The novel pharmaceutical compositions of the invention having antigonado-hypophysial, progestomimetic and progeatative activity are comprised of an effective amount of 170:,2l-dimethyl-l9-nOr-A -pregnadiene- 3,20-dione and a pharmaceutical carrier. The compositions may be in the form of injectable solutions or suspensions in ampoules or multiple dose flacons' or in the form of implants, tablets, coated tablets, sublingual tablets, capsules and suppositories. The usual individual dose of active compound for the adult is 0.1 to 2.5 mg depending upon the method of administration.

The compound of the invention is useful as progestative agent, as inhibiting hypophysis agent with predominance of anti-LH, and as antiestrogenic agent. It may be used alone or in association with an estrogen such as cthynylestradiol, mestranol or llB-methoxy-ethynyl estradiol in an estrcprogestative formulation with contraceptive action. The compositions of the invention may be used for the treatment of prostatic adenoma,

preferred embodiments to illustrate the invention.

However, it is to be understood that the inventionis not intended to be limited to the specific embodiments.

EXAMPLE PREPARATION OF pregnadiene-3,20-dione STEP A: 3-methoxy-17a-methyl-l9-nor-A- pregnatriene-ZO-one 1.150 goflithium were added with stirring to ,1 liter of ammonia cooled to C under an inert atmosphere, and after stirring for 15 minutes, 1 liter of ether .was added while maintaining the temperature at about C followed by the addition of .20 g of 3-methoxy- 19nor-A" -pregnate traene-20-one. The reaction 1701,21 dimethyl-l9-nor-A mixture was left for 2 hours with stirring at 75C under an inert atmosphere andthen 160 ccof methyl iodide were added to the reaction mixture. After ,stir ring for another 2 hours at 7'5C, the ammonia was distilled off and the residue wasadded to 1 liter of water. The aqueous phase was extractedwith etherand the combined ether phases were washed with water until the wash waterswere neutral, were dried over sodium sulfate, filtered andtdistilled to dryness. The 21 g of residue .wasdissolved in 210 cc of refluxing ethanol and after the addition of 21 cc ofv acetic acidand 21g of Girards reagent T, the mixturewas-refluxed .with stirring for 1% hours under an inert atmosphere. The

reaction mixture was then cooledto room temperature and-was added to 1,050 cc of water. cc of 2N sodium hydroxide were added thereto and the mixture was extracted with ether, the combined ether phases were washed with water until the wash waters were neutral, then weredried over sodium sulfate, filtered and evaporated to dryness to obtain 16.80 g ofraw product. The latter was purified-by dissolution in refluxing acetone and crystallization-from hotand cold acetone to obtain 3-rnethoxy-l7a-methyl-19-nor- A -pregnatriene-20-one.

STEP B: 3-methoxy-17a-methyl-l9-nor-a -pregnadiene-ZO-ol 1 A solution of 20 g of 3-methoxy-l7a-methy1-19-nor- A -pregnatriene-20-one in 400 cc of tetrahydrofuran was added to 500 cc of ammonia and then 10 cc of ethanol'were added. The interior temperature was lowered to about 35C and then 2.15 g of lithium were added'thereto under an inert atmosphere. After agitating for 15 minutes, 10 cc of ethanol and another 2.15 g of lithium were added thereto. The mixture .was stirred for 15 minutes and again 30 cc of ethanol and then 2.1 5 g of lithium were added. After 30 minutes at 35C, 30 cc of ethanol were added and the ammonia was evaporated with the temperature returning to 20C. 500 cc of water were added thereto and the mixture was extracted with ether. After the extraction with ether, the aqueous phase was drawn off and the combined ether phases were washed with water, dried over sodium sulfate, filtered and distilled to dryness to obtain 3-methoxy-l7a-methyl-l9-nor-A -pregnadiene-ZO-ol which was used as is for the next step.

STEP C: l7a-rnethyl-19nor-A -pregnene-20-ol- 3-one A solution of 20 g of next step. STEP D: 17a-methyl-19-nor-N" -pregnene-3 ,20- dione 21 cc of a dilute solution containing 5.7 g of chromic anhydride and'4.8 cc of concentrated sulfuric acid were added with agitation and under a nitrogen atmosphere to a solution of 20.5 g of 17a-methyl-19-nor- 'A -pregnene-ZO-OI-3-one in 615 cc of acetone cooled to 20C and the mixture was allowed to stand at 10C for l hour and was then poured in 2 liters of a water-ice mixture. The mixture was extracted with benzene and the combined extracts were washed first with water, then a saturated sodium bicarbonate solution and then with water. The extracts were dried over magnesium sulfate and then were distilled to dryness to obtain 20.40 g of raw product. The latter was purified by chromatography over magnesium silicate, elution with benzene containing 2.5 percent of acetone and crystallization from isopropyl ether to obtain 17amethyl- 1 9-nor-'A"-pregnene-3 ,20-dione.

STEP E: 1701,21dimethyl-l9-nor-A"-pregnadiene- 3,20-dione 16.3 cc of a solution of 29 percent of bromine in methanol were added with agitation under a nitrogen atmosphere to a solution of 8.50 g of l7a-methyl-l9- nor-'A -pregnene-B,20-dione in 85 cc of pyridine cooled to 0C and the mixture was stirred for 30 minutes at 0C. The temperature was allowed to return to room temperature and the mixture was stirred for 16 hours. The mixture was added to 850 cc of a water-ice mixture and 82 cc of hydrochloric acid were added thereto. The mixture was extracted with methylene chloride and the combined extracts were washed with water until the wash waters were neutral, were dried over magnesium sulfate and distilled to dryness to obtain 8.480 g of crude product which is purified by cristallization from isopropyl ether to obtain 5.810 g of 17- a=methyl-l9-nor-A -pregnadiene-3,2O-dione melting at 106C and identical to the product of Belgian patent No. 674,178.

The mother liquors from the purification of the said product were combined and evaporated to dryness. The residue was fractionated by chromatography over silica gel (Kieselgcl) and elution with a 7-3 mixture of benzene-ethyl acetate. The first fractions were discarded and the ensuing fraction was evaporated to obtain colorless crystals. The product was purified by empasting with 5 volumes of boiling isopropyl ether and the crystals formed after cooling were recovered by vacuum filtration, were washed twice with 2-volumes of isopropyl ether and dried in a ventilated atmosphere to obtain l7a,2ldimethyl-19-nor-N- -pregnadiene- 3,20-dione melting at 152C and having a specific rotation [owr 262 (c 0.5% in ethanol). The creamwhite product was soluble in acetone and benzene and insoluble in water. Analysis: C H O molecular weight 326.45 Calculated: C 80.93 H 9.26 Found: 80.6-80.7 9.2-9.3 U. V. Spectrum (ethanol): Max. at 215 mu E 202 lnflex towards 234 mu E 155 lnflex. towards 245 mp. 15 120 'r max. at 305 mp. E 648 or e 21,150 RMN Spectrum in accord with proposed structure 2l-methyl-at 55, 6L5 and 68 Hz l7-methyl-at 66Hz 4-ethylenic proton-3 39 l3-methyl-at 46.5 Hz

As far as is known, this compound is not described in the literature.

PHARMACOLOGICAL STUDY A. Progestomimetic Activity The progestomimetic activity was determined by the Clauberg test using immature rabbits previously sensitized 5 days earlier by subcutaneously administering 10 pg of estradiol benzoate. They are treated with doses of the test compound for 5 days and are killed on the sixth day. Sections of the uterus are examined in Mac- Phail units for the proliferation of endometric lacework, characteristic of progestomimetic action. The test compound was administered subcutaneously as a solution in olive oil containing 5 percent benzylic alcohol at a dose of 1.56, 3.12 and 6.25 y. The results are reported in Table I.

TABLE 1 Product Doses in MacPhail 'y/day Units :,2 l dirnethyll 9-nor-A"-pregna-diene-3,20-dione 1.56 2.0 3.1 2.6 6.2 2.7

The results of Table 1 show that the test compound has a clear progestomimetic activity at a dose of 1.567. Under similar test conditions, fi-chloro-lia-acetoxy- 'A regnadiene-B,20-dione at 1.56 7 per day had a MacPhail of 1.0, at a dose of 3.127 an index of 2.4. Therefore the compound of the invention has twice the progestomimetic activity of 6-chloro-l7a-acetoxy-b pregnadiene-3,20-dione when administered subcutaneously.

B. Antigonadotrophic Activity The antigonadotrophic activity was determined on puberic rats weighing about 200 g. The compounds were administered subcutaneously as a solution in seaame oil containing 5 percent benzylic alcohol in a volume of 0.2 cc in 12 treatments over 14 days at the daily dose of 0.2 and 1 mg per rat. On the 15th day, the rats were sacrificed by carotidienic bleeding and the seminal vesicles, prostate, testicles and surrenals were removed and weighed. The results are reported in Table 11.

TABLE 11 seminal testicles vesicles prostate surrenals product doses in g in mg in mg in mg control 2.04 529.6 320.2 40.3 1711,21 dimethyl- 19 ZOO-y 2.56 272.0 244.0 49.1 nova, 9- (-50%) (24%) pregnadiene-l mg 2.64 145.6 172.5 45.5

dione (-73%) (-46%) The results of Table 11 show that the compound of the invention possesses a clear, predominantly anti-LH antigo-nadotrophic activity at a close of 200 without provoking surrenalienic aplasia.

C. Antiestrogenic Activity The antiestrogenic activity was determined on immature mice by a technique inspired by the test of Rubin [Endo., Vol. 49 (1951), p.429] and similar to that of Dorfman et al. [Methods in Hormone Research, Vol. ll, 1962, p. 118]. Groups of 4 mice 19 to 21 days old received daily subcutaneously for three days an injection of estradiol alone, an injection of the test product alone or an injection of estradiol and the test product. In the last case, the two steroids were injected at different points. The animals were killed on the fourth day and the uterus was removed and weighed.

The estradiol in solution in olive oil containing percent benzylic alcohol was administered at a total dose of 0.277, each injection having a volume of 0.1 cc per mouse. The test compound in solution in olive oil containing 5 percent benzylic alcohol was administered to two lots of mice at increasing doses of 1.1 3.3 and y and also 3.3 107 and 307. The results reported below show a clear anti-estrogenic activity.

1.1-y 31 percent of the effect of 0.277 of estradiol 3.37 37 percent 10.07 47 percent 3.37 32 percent 10.07 47 percent 30.0y 38 percent D. Maintaining of Gestation Puberic rats weighing about 250 g were left in contact with male rats and the first day of gestation (J-l) was determined by the presence of spermatozoon in a vaginal smear. On the 8th day (J-8), the female rats were castrated under ether anesthesia and from the 8th to the 19th day, the test product was subcutaneously administered every day in solution in sesame oil containing 5% benzylic alcohol at a volume of 0.5 cc. On the 20th day (J-20), the animals were killed and the activity of the product was determined by the number of fetuses in term at J-20 with respect to the number of implantations at J-8 expressed as a percentage. The results are reported in Table 111.

TABLE Ill fetuses rats in per no. dose gestation placentas implantations Product in mg .l-8 .I-20 J-B .l-20 at J-8 controls 0 8 0 9.1

1701,21 A di- 2 8 8 12.0 12.0 72% methyll9-nor- 0.5 8 8 10.6 10.6 74% A1.9

pregna diene- 3,20-dione The results of Table 111 show that the compound of the invention almost totally maintained gestation in rats at a dose of 0.5 mg.

Various modifications of the compositions and method of the invention may be made without departing from the spirit or scope thereof and it is to be understood that the invention is to be limited only as defined in the appended claims.

We claim:

1. A pharmaceutical composition comprising an effective amount of 17a,2l-dimethyl-19-nor-A -pregnadiene-3,20-dione and a pharmaceutical carrier.

2. A composition of claim 1 which also contains an estrogen.

3. A method of inducing progestomimetic activity in warm-blooded animals which comprises administering to warm-blooded animals a safe and effective amount of 17a,2l-dimethyl-l9-nor-A" -pregnadiene-3,20- dione.

4. A method of inducing antigonado-hypophysial activity in warm-blooded animals which comprises administering to warm-blooded animals a safe and effective amount of 1711,21-dimethyl-19-nor-A" -pregnadiene-3,20-dione.

5. A method of inducing progestative activity in warm-blooded animals which comprises administering to warm-blooded animals a safe and effective amount of 1 701,2 l-dimethyl- 1 9-nor-A--pregnadiene-3 ,20- dione. 

2. A composition of claim 1 which also contains an estrogen.
 3. A method of inducing progestomimetic activity in warm-blooded animals which comprises administering to warm-blooded animals a safe and effective amount of 17 Alpha ,21-dimethyl-19-nor- Delta 4,9-pregnadiene-3,20-dione.
 4. A method of inducing antigonado-hypophysial activity in warm-blooded animals which comprises administering to warm-blooded animals a safe and effective amount of 17 Alpha ,21-dimethyl-19-nor- Delta 4,9-pregnadiene-3,20-dione.
 5. A method of inducing progestative activity in warm-blooded animals which comprises administering to warm-blooded animals a safe and effective amount of 17 Alpha ,21-dimethyl-19-nor- Delta 4,9-pregnadiene-3,20-dione. 